重组Anti-GIRK2抗体[EPR23841-83]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labelling GIRK2 with ab259909 at 1/200 (2.345 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab259909 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Cytoplasmic staining on human cerebrum (PMID : 8642402).

Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

IHC image of GIRK2 staining in a section of frozen human normal substantia nigra performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab259909, 0.05 μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of paraffin-embedded human insulinoma tissue labelling GIRK2 with ab259909 at 1/200 (2.345 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab259909 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Cytoplasmic staining on human insulinoma (PMID : 7592809).

Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Negative tissue control image : IHC image of GIRK2 staining in a section of frozen human normal liver performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab259909, 0.05 μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of paraffin-embedded human liver tissue labelling GIRK2 with ab259909 at 1/200 (2.345 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab259909 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Negative control : No staining on human liver (PMID : 7592809).

Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat hippocampus tissue labeling GIRK2 with ab259909 at 1/100 (4.69 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). The nuclear counterstain was DAPI (Blue).

Cytoplasmic staining on rat hippocampus (PMID : 8929423) is observed.

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labelling GIRK2 with ab259909 at 1/200 (2.345 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab259909 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Postive staining predominantly on granular layer of rat cerebellum (PMID : 8642402, PMID : 9023358).

Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus tissue labeling GIRK2 with ab259909 at 1/100 (4.69 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). The nuclear counterstain was DAPI (Blue).

Cytoplasmic staining on mouse hippocampus (PMID : 8929423) is observed.

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver tissue labeling GIRK2 with ab259909 at 1/100 (4.69 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). The nuclear counterstain was DAPI (Blue).

Negative control : No staining on rat liver (PMID : 7592809) is observed.

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebellum tissue labeling GIRK2 with ab259909 at 1/100 (4.69 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). The nuclear counterstain was DAPI (Blue).

Cytoplasmic staining on rat cerebellum (PMID : 8929423) is observed.

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebral cortex tissue labeling GIRK2 with ab259909 at 1/100 (4.69 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). The nuclear counterstain was DAPI (Blue).

Cytoplasmic staining on rat cerebral cortex (PMID : 8929423) is observed.

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling GIRK2 with ab259909 at 1/100 (4.69 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). The nuclear counterstain was DAPI (Blue).

Negative control : No staining on mouse liver (PMID : 7592809) is observed.

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebellum tissue labeling GIRK2 with ab259909 at 1/100 (4.69 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). The nuclear counterstain was DAPI (Blue).

Cytoplasmic staining on mouse cerebellum (PMID : 8929423) is observed.

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebral cortex tissue labeling GIRK2 with ab259909 at 1/100 (4.69 μg/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). The nuclear counterstain was DAPI (Blue).

Cytoplasmic staining on mouse cerebral cortex (PMID : 8929423) is observed.

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labelling GIRK2 with ab259909 at 1/200 (2.345 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab259909 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Postive staining predominantly on granular layer of mouse cerebellum (PMID : 8642402, PMID : 9023358 ).

Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

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Immunoprecipitation - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

GIRK2 was immunoprecipitated from 0.35 mg mouse brain tissue lysate 10 μg with ab259909 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab259909 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate 10 μg.

Lane 2 : ab259909 IP in mouse brain tissue lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab259909 in mouse brain tissue lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 23 seconds.

Input sample is non-boiled as boiling may cause protein aggregates.The expression of truncated GIRK2 proteins observed around 40KD is consistent with what has been described in the literature (PMID : 28951616).

All lanes:

Immunoprecipitation - Anti-GIRK2 antibody [EPR23841-83] (ab259909)

Predicted band size: 48 kDa

Observed band size: 40 kDa,48 kDa

false

• WB

Lab

Western blot - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Molecular weight around 40kDa, cleaved forms. (PMID : 28951616).

Samples are non-boiled as boiling may cause protein aggregates.

Negative control : liver (PMID : 8929423).

Exposure time : 5.5 seconds.

All lanes:

Western blot - Anti-GIRK2 antibody [EPR23841-83] (ab259909) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 48 kDa

Observed band size: 35 kDa,40 kDa,48 kDa

false

• WB

Lab

Western blot - Anti-GIRK2 antibody [EPR23841-83] (AB259909)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Samples are non-boiled as boiling may cause protein aggregates.

Negative control : liver (PMID : 8929423).

Exposure time : 5.5 secs.

All lanes:

Western blot - Anti-GIRK2 antibody [EPR23841-83] (ab259909) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 40 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Lane 4:

Rat liver tissue lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 48 kDa

false